Characterization and cytocompatibility of thermosensitive hydrogel embedded with chitosan nanoparticles for delivery of bone morphogenetic protein-2 plasmid DNA
Characterization and cytocompatibility of thermosensitive hydrogel embedded with chitosan nanoparticles for delivery of bone morphogenetic protein-2 plasmid DNA
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嵌入壳聚糖纳米粒子的热敏水凝胶的表征和细胞相容性，用于递送骨形态发生蛋白-2质粒DNA
DOI：
10.1007/s10856-016-5743-0
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发表时间：
2016-08-01
影响因子：
3.7
通讯作者：
Wang, Lei
中科院分区：
文献类型：
--
作者：
Li, Dan-Dan;Pan, Jian-Feng;Wang, Lei
一种新型可注射壳聚糖温敏水凝胶被设计为用于牙周组织内源性修复的靶向多效支架。该水凝胶复合物是通过将负载骨形态发生蛋白 - 2质粒DNA（pDNA - BMP2）的壳聚糖纳米粒子（CSn）嵌入含α，β - 甘油磷酸酯（α，β - GP）的壳聚糖（CS）基水凝胶中而设计的，称为CS/CSn（pDNA - BMP2）- GP。随后对其进行了表征、pDNA - BMP2的体外释放曲线以及对人牙周膜细胞（HPDLCs）的细胞相容性研究。CSn（pDNA - BMP2）的平均直径为270.1 nm，多分散指数（PDI）为0.486，ζ电位为 +27.0 mV。DNA酶I保护试验表明，CSn可保护pDNA - BMP2免受核酸酶降解。CSn（pDNA - BMP2）的包封率和载药量分别超过80%和30%。当将CSn（pDNA - BMP2）加入CS/α，β - GP体系时，溶胶 - 凝胶转变时间仅为3分钟。扫描电子显微镜显示，CSn（pDNA - BMP2）随机分散在具有规则孔洞和多孔结构的网络中。称重法显示，在pH 4.0的介质中，溶胀率和降解速度比pH 6.8时更快。体外pDNA - BMP2释放试验表明，在相同的释放介质中，pDNA - BMP2从CS/CSn - GP中的累积释放速率比从CSn中慢得多。在不同pH的释放介质中，pDNA - BMP2在pH 6.8时的释放比在pH 4.0时慢得多。与HPDLCs的三维培养显示出良好的细胞增殖，并且细胞计数试剂盒 - 8检测表明，在CS/α，β - GP中添加CSn（pDNA - BMP2）可促进细胞生长。总之，CS/CSn（pDNA - BMP2）- GP复合体系表现出优异的生物学特性和细胞相容性，表明其作为基因传递载体和牙周组织内源性修复的组织再生支架具有巨大潜力。
A novel injectable chitosan thermosensitive hydrogel was designed as a target multi-effect scaffold for endogenous repair of the periodontium. The hydrogel complex was designed by embedding chitosan nanoparticles (CSn) loaded with bone morphogenetic protein-2 plasmid DNA (pDNA-BMP2) into a chitosan (CS)-based hydrogel with α,β-glycerophosphate (α,β-GP), termed CS/CSn(pDNA-BMP2)-GP. Characterization, the in vitro release profile for pDNA-BMP2, and cytocompatibility to human periodontal ligament cells (HPDLCs), were then conducted. The average diameter of the CSn(pDNA-BMP2) was 270.1 nm with a polydispersity index (PDI) of 0.486 and zeta potential of +27.0 mv. A DNase I protection assay showed that CSn could protect the pDNA-BMP2 from nuclease degradation. Encapsulation efficiency and loading capacity of CSn(pDNA-BMP2) were more than 80 and 30 %, respectively. The sol–gel transition time was only 3 min when CSn(pDNA-BMP2) was added into the CS/α,β-GP system. Scanning electron microscopy showed that CSn(pDNA-BMP2) was randomly dispersed in a network with regular holes and a porous structure. Weighting method showed the swelling ratio and degradation was faster in medium of pH 4.0 than pH 6.8. An in vitro pDNA-BMP2 release test showed that the cumulative release rate of pDNA-BMP2 was much slower from CS/CSn-GP than from CSn in identical release media. In release media with different pH, pDNA-BMP2 release was much slower at pH 6.8 than at pH 4.0. Three-dimensional culture with HPDLCs showed good cell proliferation and the Cell-Counting Kit-8 assay indicated improved cell growth with the addition of CSn(pDNA-BMP2) to CS/α,β-GP. In summary, the CS/CSn(pDNA-BMP2)-GP complex system exhibited excellent biological properties and cytocompatibility, indicating great potential as a gene delivery carrier and tissue regeneration scaffold for endogenous repair of the periodontium.